Garden bean variety PV-803

ABSTRACT

A novel garden bean variety, designated PV-803, is disclosed. The invention relates to the seeds of garden bean variety PV-803, to the plants of garden bean line PV-803 and to methods for producing a bean plant by crossing the variety PV-803 with itself or another bean line. The invention further relates to methods for producing a bean plant containing in its genetic material one or more transgenes and to the transgenic plants produced by that method and to methods for producing other garden bean lines derived from the variety PV-803.

BACKGROUND OF THE INVENTION

The present invention relates to a new and distinctive garden beanvariety (Phaseolus vulgaris), designated PV-803. All publications citedin this application are herein incorporated by reference.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possesses the traits tomeet the program goals. The goal is to combine in a single variety animproved combination of desirable traits from the parental germplasm.

In beans, these important traits may include fresh pod yield, higherseed yield, resistance to diseases and insects, better stems and roots,tolerance to drought and heat, and better agronomic quality. Withmechanical harvesting of many crops, uniformity of plant characteristicssuch as germination and stand establishment, growth rate, maturity andplant height is important.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar will incur additional coststo the seed producer, the grower, processor and consumer for specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

Garden bean, Phaseolus vulgaris L., is an important and valuablevegetable crop. Thus, a continuing goal of garden bean plant breeders isto develop stable, high yielding garden bean cultivars that areagronomically sound. The reasons for this goal are obviously to maximizethe amount of yield produced on the land. To accomplish this goal, thegarden bean breeder must select and develop garden bean plants that havetraits that result in superior cultivars.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools and methods which are meant to beexemplary, not limiting in scope. In various embodiments, one or more ofthe above-described problems have been reduced or eliminated, whileother embodiments are directed to other improvements.

According to the invention, there is provided a novel garden beanvariety designated PV-803. This invention thus relates to the seeds ofgarden bean variety PV-803, to the plants or part(s) thereof of gardenbean variety PV-803, to plants or part(s) thereof having all thephenotypic and morphological characteristics of garden bean varietyPV-803 and to plant or part(s) thereof having the phenotypic andmorphological characteristics of garden bean variety PV-803 listed inTable 1 as determined at the 5% significance level when grown in thesame environmental conditions. Plant parts of the garden bean variety ofthe present invention are also provided such as, i.e., pollen obtainedfrom the plant variety and an ovule obtained from the plant variety.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of bean variety PV-803. The tissue culture willpreferably be capable of regenerating plants having the physiologicaland morphological characteristics of bean variety PV-803. Preferably,the cells of such tissue culture will be embryos, meristematic cells,seeds, callus, pollen, leaves, anthers, pistils, roots, root tips, pods,flowers and stems. Protoplasts produced from such tissue culture arealso included in the present invention. The bean plants regenerated fromthe tissue culture are also part of the invention.

Also included in the invention are methods for producing a bean plantproduced by crossing bean variety PV-803 with itself or another beanvariety. When crossed with itself, i.e., when crossed with another beanvariety PV-803 plant or self-pollinated, bean variety PV-803 will beconserved. When crossed with another, different bean plant, an F₁ hybridseed is produced. F₁ hybrid seeds and plants produced by growing saidhybrid seeds are included in the present invention. A method forproducing an F₁ hybrid bean seed comprising crossing a bean varietyPV-803 plant with a different bean plant and harvesting the resultanthybrid bean seed are also part of the invention. The hybrid bean seedproduced by the method comprising crossing a bean variety PV-803 plantwith a different bean plant and harvesting the resultant hybrid beanseed, are included in the invention, as are the hybrid bean plant orpart(s) thereof, and seeds produced by growing said hybrid bean seed.

In another aspect, the present invention provides transformed PV-803bean variety plants or part(s) thereof that have been transformed sothat its genetic material contains one or more transgenes, preferablyoperably linked to one or more regulatory elements. Also, the inventionprovides methods for producing a bean plant containing in its geneticmaterial one or more transgenes, preferably operably linked to one ormore regulatory elements, by crossing transformed PV-803 bean varietyplants with either a second plant of another bean variety, or anon-transformed PV-803 bean variety, so that the genetic material of theprogeny that results from the cross contains the transgene(s),preferably operably linked to one or more regulatory elements. Theinvention also provides methods for producing a bean plant that containsin its genetic material one or more transgene(s), wherein the methodcomprises crossing the variety PV-803 with a second bean variety ofanother bean variety which contains one or more transgene(s) operablylinked to one or more regulatory element(s) so that the genetic materialof the progeny that results from the cross contains the transgene(s)operably linked to one or more regulatory element(s). Transgenic beancultivars, or part(s) thereof produced by the methods are in the scopeof the present invention.

More specifically, the invention comprises methods for producing a malesterile bean plant, an herbicide resistant bean plant, an insectresistant bean plant, a disease resistant bean plant, a water stresstolerant bean plant, a heat stress tolerant bean plant, and a bean plantwith improved shelf-life. Said methods comprise transforming a beanvariety PV-803 plant with a nucleic acid molecule that confers malesterility, herbicide resistance, insect resistance, disease resistance,water stress tolerance, heat stress tolerance, or improved shelf life,respectively. The transformed bean plants, or part(s) thereof, obtainedfrom the provided methods, including a male sterile bean plant, anherbicide resistant bean plant, an insect resistant bean plant, adisease resistant bean plant, a bean plant tolerant to water stress, abean plant tolerant to heat stress or a bean plant with improvedshelf-life are included in the present invention. For the presentinvention and the skilled artisan, disease is understood to be fungaldiseases, viral diseases, bacterial diseases or other plant pathogenicdiseases and a disease resistant plant will encompass a plant resistantto fungal, viral, bacterial and other plant pathogens.

In another aspect, the present invention provides for methods ofintroducing one or more desired trait(s) into bean variety PV-803 andplants obtained from such methods. The desired trait(s) may be, but notexclusively, a single gene, preferably a dominant but also a recessiveallele. Preferably, the transferred gene or genes will confer suchtraits as male sterility, herbicide resistance, insect resistance,resistance to bacterial, fungal, or viral disease, increased leafnumber, improved shelf-life, and tolerance to water stress or heatstress. The gene or genes may be naturally occurring gene(s) ortransgene(s) introduced through genetic engineering techniques. Themethod for introducing the desired trait(s) is preferably a backcrossingprocess making use of a series of backcrosses to bean variety PV-803during which the desired trait(s) is maintained by selection.

In a preferred embodiment, the present invention provides methods forincreasing and producing bean variety PV-803 seed, whether by crossing afirst parent bean variety plant with a second parent bean variety plantand harvesting the resultant bean seed, wherein both said first andsecond parent bean variety plant are the bean variety PV-803 or byplanting a bean seed of the bean variety PV-803, growing a bean varietyPV-803 plant from said seed, controlling a self pollination of the plantwhere the pollen produced by a grown bean variety PV-803 plantpollinates the ovules produced by the very same bean variety PV-803grown plant, and harvesting the resultant seed.

The invention further provides methods for developing bean cultivars ina bean breeding program using plant breeding techniques includingrecurrent selection, backcrossing, pedigree breeding, molecular markers(Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms(RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily PrimedPolymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting(DAF), Sequence Characterized Amplified Regions (SCARs). AmplifiedFragment Length Polymorphisms (AFLPs), and Simple Sequence Repeats(SSRs) which are also referred to as Microsatellites, etc.) enhancedselection, genetic marker enhanced selection, and transformation. Seeds,bean plants, and part(s) thereof produced by such breeding methods arealso part of the invention.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DETAILED DESCRIPTION OF THE INVENTION

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele. An allele is any of one or more alternative forms of a genewhich relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Anthracnose. A seedling disease caused by the pathogen Colletotrichumlindemuthianum L. Symptoms include dark brown to black sunken lesions onthe cotyledons and stems. Severely infected cotyledons senesceprematurely, and growth of the plant is stunted. Diseased areas maygirdle the stem and kill the seedling. Also known as Fungus BeanAnthracnose.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotypes of the F₁hybrid.

Bacterial Brown Spot. A bean disease caused by the pathogen Pseudomonassyringae pv. Syringae. Symptoms include small water-soaked spots thatdevelop into distinctive necrotic brown spots about 3-8 mm in diameteron the pods, often with a narrow, diffuse yellow margin.

Bean Common Mosaic Virus (BCMV). An insect transmitted disease thatoccurs worldwide causing low quality harvest product and potentialoverall crop reduction in yield from 80% to 100%. Resistance to thisdisease is highly desirable.

Bean Rust. A defoliating bean disease caused by the pathogen Uromycesappendiculatus. Symptoms include rust-colored pustules often surroundedby a chlorotic halo. The disease occurs worldwide and can destroy anentire crop if conditions are favorable.

Bean yield (tons/acre). The yield in tons/acre is the actual yield ofthe bean pods at harvest.

Determinate plant. A determinate plant will grow to a fixed number ofnodes while an indeterminate plant continues to grow during the season.

Emergence. The rate that the seed germinates and sprouts out of theground.

Essentially all the physiological and morphological characteristics. Aplant having essentially all the physiological and morphologicalcharacteristics means a plant having the physiological and morphologicalcharacteristics, except for the characteristics derived from theconverted gene.

Field holding ability. A bean plant that has field holding ability meansa plant having pods that remain smooth and retain their color even afterthe seed is almost fully developed.

Immunity to disease(s) and or insect(s). A bean plant which is notsubject to attack or infection by specific disease(s) and or insect(s)is considered immune.

Intermediate resistance to disease(s) and or insect(s). A bean plantthat restricts the growth and development of specific disease(s) and orinsect(s), but may exhibit a greater range of symptoms or damagecompared to resistant plants. Intermediate resistant plants will usuallyshow less severe symptoms or damage than susceptible plant varietieswhen grown under similar environmental conditions and/or specificdisease(s) and or insect(s) pressure, but may have heavy damage underheavy pressure. Intermediate resistant bean plants are not immune to thedisease(s) and or insect(s).

Machine harvestable bush. A machine harvestable bush means a bean plantthat stands with pods off the ground. The pods can be removed by amachine from the plant without leaves and other plant parts.

Maturity. A maturity under 53 days is considered early while maturitybetween 54-59 days is considered average or medium and maturity of 60 ormore days would be late.

Maturity date. Plants are considered mature when the pods have reachedtheir maximum allowable seed size and sieve size for the specific useintended. This can vary for each end user, e.g., processing at differentstages of maturity would be required for different types of consumerbeans, such as “whole pack,” “cut,” or “french style.” The number ofdays are calculated from a relative planting date which depends on daylength, heat units, and other environmental factors.

Plant adaptability. A plant having good plant adaptability means a plantthat will perform well in different growing conditions and seasons.

Plant architecture. Plant architecture is the shape of the overall plantwhich can be tall-narrow, short-wide, medium height, and/or mediumwidth.

Plant cell. As used herein, the term “plant cell” includes plant cellswhether isolated, in tissue culture, or incorporated in a plant or plantpart.

Plant habit. A plant can be erect (upright) to sprawling on the ground.

Plant height. Plant height is taken from the top of the soil to the topnode of the plant and is measured in centimeters or inches.

Plant part. As used herein, the term “plant part” includes leaves,stems, roots, seed, embryos, pollen, ovules, flowers, root tips,anthers, tissue, cells, pods, and the like.

Pod set height. The pod set height is the location of the pods withinthe plant. The pods can be high (near the top), low (near the bottom),or medium (in the middle) of the plant.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Resistance to disease(s) and or insect(s). A bean plant that restrictsthe growth and development of specific disease(s) and or insect(s) undernormal disease(s) and or insect(s) attack pressure when compared tosusceptible plants. These bean plants can exhibit some symptoms ordamage under heavy disease(s) and or insect(s) pressure. Resistant beanplants are not immune to the disease(s) and or insect(s).

RHS. RHS refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Hort. Society Enterprise Ltd. RHS Garden; Wisley,Woking, Surrey GU236QB, UK.

Sieve size (sv). Sieve size 1 means pods that fall through a sievegrader which culls out pod diameters of 4.76 mm through 5.76 mm. Sievesize 2 means pods that fall through a sieve grader which culls out poddiameters of 5.76 mm through 7.34 mm. Sieve size 3 means pods that fallthrough a sieve grader which culls out pod diameters of 7.34 mm through8.34 mm. Sieve size 4 means pods that fall through a sieve grader whichculls out pod diameters of 8.34 mm through 9.53 mm. Sieve size 5 meanspods that fall through a sieve grader which culls out pod diameters of9.53 mm through 10.72 mm. Sieve size 6 means pods that fall through asieve grader that will cull out pod diameters of 10.72 mm or larger.

Single gene converted (conversion). Single gene converted (conversion)plants refers to plants which are developed by a plant breedingtechnique called backcrossing wherein essentially all of the desiredmorphological and physiological characteristics of a variety arerecovered in addition to the single gene transferred into the varietyvia the backcrossing technique or via genetic engineering.

Slow seed development. Beans having slow seed development develop seedslowly even after the pods are full sized. This characteristic gives tothe cultivar its field holding ability.

Susceptible to disease(s) and or insect(s). A bean plant that issusceptible to disease(s) and or insect(s) is defined as a bean plantthat has the inability to restrict the growth and development ofspecific disease(s) and or insect(s). Plants that are susceptible willshow damage when infected and are more likely to have heavy damage undermoderate levels of specific disease(s) and or insect(s).

Uromyces. Also referred to as ‘bean rust,’ is a defoliating bean diseasecaused by the pathogen Uromyces appendiculatus. Symptoms includerust-colored pustules often surrounded by a chlorotic halo. The diseaseoccurs worldwide and can destroy an entire crop if conditions arefavorable.

Garden bean variety PV-803 most closely resembles garden bean varietyValentino (U.S. Plant Variety Protection No. 200600097). The comparativecharacteristic that most readily distinguishes the two varietiesincludes, but may not be limited to, reaction to fungus beanAnthracnose. Valentino is susceptible to fungus bean AnthracnoseColletotrichum lindemuthianum race Lambda, whereas PV-803 is highlyresistant to fungus bean Anthracnose. Multiple seedling tests over thepast few years have shown that Valentino dies in less than 10 days afterinoculation with fungus bean Anthracnose, while PV-803 shows nosusceptibility symptoms of fungus bean Anthracnose.

Additionally, garden bean variety PV-803 has slightly darker and thickerpods than Valentino. Some of the selection criteria used for variousgenerations include: pod appearance, pod length, pod diameter, beanyield, pod set height, emergence, maturity, plant architecture, habitand height, seed yield, and quality and disease resistances.

Garden bean variety PV-803 has shown uniformity and stability for thetraits, as described in the following Variety Description Information.It has been self-pollinated a sufficient number of generations withcareful attention to uniformity of plant type. The variety has beenincreased with continued observation for uniformity. No variant traitshave been observed or are expected in garden bean variety PV-803.

Garden bean variety PV-803 has has the following morphologic and othercharacteristics (based primarily on data collected at Belle Glade, Fla.and Andijk, The Netherlands).

TABLE 1 VARIETY DESCRIPTION INFORMATION Market Maturity: Days to ediblepods: 56 Number of days later than ‘Pike’ (U.S. Pat. No. 7,511,197): 1Plant: Habit: Determinate Height: 50.0 cm Spread: 50.0 cm Pod position:Medium-high Bush form: High bush Leaves: Surface: Strong rugosity Size:Medium to large Color: Dark green Anthocyanin Pigment: Flowers: AbsentStems: Absent Pods: Absent Seeds: Absent Leaves: Absent Petioles: AbsentPeduncles: Absent Nodes: Absent Flower Color: Color of standard: WhiteColor of wings: White Color of keel: White Pods (edible maturity):Exterior color: Very dark green Cross section pod shape: RoundCreaseback: Absent Pubescence: Sparse Constriction: Slight Spur length:Approximately 16.0 mm Fiber: Sparse Number of seeds per pod: Average 5Suture string: Absent Seed development: Medium Machine harvest: AdaptedDistribution of sieve size at optimum maturity: 80% Sieve 4 20% Sieve 5Seed Color: Seed coat luster: Shiny Primary color: White Seed coatpattern: Solid Hilar ring: Present; slightly yellow Seed Shape and Size:Hilum view: Elliptical Cross section: Broad elliptic Side view: ReniformSeed size (g/100 seeds): Approximately 26 g Disease Resistance: BeanCommon Mosaic Virus (BCMV I gene): Resistant; I-gene present Pseudomonassyringae pv. syringae (Bacterial Brown Spot): Intermediate- susceptibleColletotrichum lindemuthianum L. (Anthracnose): Resistant to race LambdaUromyces appendiculatus (Bean rust) Resistant to races 41 and 49

Further Embodiments of the Invention

This invention also is directed to methods for producing a garden plantby crossing a first parent bean plant with a second parent bean plantwherein either the first or second parent bean plant is a bean plant ofthe line PV-803. Further, both first and second parent bean plants cancome from variety PV-803. When self pollinated, or crossed with anotherbean variety PV-803 plant, the bean variety PV-803 will be stable, whilewhen crossed with another, different bean variety plant, an F₁ hybridseed is produced.

Still further, this invention also is directed to methods for producingan PV-803-derived bean plant by crossing variety PV-803 with a secondbean plant and growing the progeny seed, and repeating the crossing andgrowing steps with the variety PV-803-derived plant from 0 to 7 times.Thus, any such methods using the variety PV-803 are part of thisinvention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using variety PV-803 as aparent are within the scope of this invention, including plants derivedfrom variety PV-803. Advantageously, the variety is used in crosses withother, different, cultivars to produce first generation (F₁) bean seedsand plants with superior characteristics.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, recurrent selection andbackcross breeding.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant cultivars.Nevertheless, it is also suitable for the adjustment and selection ofmorphological character, color characteristics and simply inheritedquantitative characters. Various recurrent selection techniques are usedto improve quantitatively inherited traits controlled by numerous genes.The use of recurrent selection in self-pollinating crops depends on theease of pollination, the frequency of successful hybrids from eachpollination and the number of hybrid offspring from each successfulcross.

Each breeding program should include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended, etc.).

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for three or more years. The best lines are candidatesfor new commercial cultivars. Those still deficient in a few traits maybe used as parents to produce new populations for further selection.

These processes, which lead to the final step of marketing anddistribution, usually take from eight to twelve years from the time thefirst cross is made. Therefore, development of new cultivars is atime-consuming process that requires precise forward planning, efficientuse of resources, and a minimum of changes in direction.

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

The goal of garden bean plant breeding is to develop new, unique andsuperior garden bean cultivars. The breeder initially selects andcrosses two or more parental lines, followed by repeated selfing andselection, producing many new genetic combinations. The breeder cantheoretically generate billions of different genetic combinations viacrossing, selfing and mutations.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under unique and differentgeographical, climatic and soil conditions and further selections arethen made during and at the end of the growing season. The cultivarsthat are developed are unpredictable. This unpredictability is becausethe breeder's selection occurs in unique environments with no control atthe DNA level (using conventional breeding procedures), and withmillions of different possible genetic combinations being generated. Abreeder of ordinary skill in the art cannot predict the final resultinglines he develops, except possibly in a very gross and general fashion.This unpredictability results in the expenditure of large amounts ofresearch monies to develop superior new garden bean cultivars.

The development of new garden bean cultivars requires the developmentand selection of garden bean varieties, the crossing of these varietiesand the evaluation of the crosses.

Pedigree breeding and recurrent selection breeding methods are used todevelop cultivars from breeding populations. Breeding programs combinedesirable traits from two or more cultivars or various broad-basedsources into breeding pools from which cultivars are developed byselfing and selection of desired phenotypes.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents that possess favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁s or by intercrossing two F₁s (sib mating). Selection of the bestindividuals may begin in the F₂ population; then, beginning in the F₃,the best individuals in the best families are selected. Replicatedtesting of families, or hybrid combinations involving individuals ofthese families, often follows in the F₄ generation to improve theeffectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified, or created,by intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror inbred line which is the recurrent parent. The source of the trait tobe transferred is called the donor parent. The resulting plant isexpected to have the attributes of the recurrent parent (e.g., cultivar)and the desirable trait transferred from the donor parent. After theinitial cross, individuals possessing the phenotype of the donor parentare selected and repeatedly crossed (backcrossed) to the recurrentparent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In a multiple-seed procedure, garden bean breeders commonly harvest oneor more pods from each plant in a population and thresh them together toform a bulk. Part of the bulk is used to plant the next generation andpart is put in reserve. The procedure has been referred to as modifiedsingle-seed descent or the pod-bulk technique.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster to thresh pods with a machine than to remove oneseed from each by hand for the single-seed procedure. The multiple-seedprocedure also makes it possible to plant the same number of seeds of apopulation each generation of inbreeding. Enough seeds are harvested tomake up for those plants that did not germinate or produce seed.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., Principles of Plant Breeding, John Wiley and Son, pp.115-161, 1960; Allard, 1960; Fehr, 1987)

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which garden bean plantscan be regenerated, plant calli, plant clumps and plant cells that areintact in plants or parts of plants, such as embryos, pollen, ovules,flowers, seeds, pods, stems, roots, anthers, pistils, root tips, leaves,and the like.

As is well known in the art, tissue culture of garden bean can be usedfor the in vitro regeneration of a garden bean plant. Tissue culture ofvarious tissues of garden beans and regeneration of plants therefrom iswell known and widely published. For example, reference may be had toMcClean, P., Grafton, K. F., “Regeneration of dry bean (Phaseolusvulgaris) via organogenesis,” Plant Sci., 60, 117-122 (1989); Mergeai,G., Baudoin, J. P., “Development of an in vitro culture method forheart-shaped embryo in Phaseolus vulgaris,” B.I.C. Invit. Papers 33,115-116 (1990); Vanderwesthuizen, A. J., Groenewald, E. G., “RootFormation and Attempts to Establish Morphogenesis in Callus Tissues ofBeans (Phaseolus-vulgaris L.),” S. Afr. J. Bot. 56, 271-273 (2 Apr.1990); Benedicic, D., et al., “The regeneration of Phaseolus vulgaris L.plants from meristem culture,” Abst. 5th I.A.P.T.C. Cong. 1, 91 (#A3-33)(1990); Genga, A., Allavena, A., “Factors affecting morphogenesis fromimmature cotyledons of Phaseolus coccineus L.,” Abst. 5th I.A.P.T.C.Cong. 1, 101 (#A3-75) (1990); Vaquero, F., et al., “Plant regenerationand preliminary studies on transformation of Phaseolus coccineus,” Abst.5th I.A.P.T.C. Cong. 1, 106 (#A3-93) (1990); Franklin, C. I., et al.,“Plant Regeneration from Seedling Explants of Green Bean(Phaseolus-Vulgaris L.) via Organogenesis,” Plant Cell Tissue Org.Cult., 24, 199-206 (3 Mar. 1991); Malik, K. A., Saxena, P. K.,“Regeneration in Phaseolus-vulgaris L.—Promotive Role ofN6-Benzylaminopurine in Cultures from Juvenile Leaves,” Planta, 184(1),148-150 (1991); Genga, A., Allavena, A., “Factors affectingmorphogenesis from immature cotyledons of Phaseolus coccineus L.,” PlantCell Tissue Org. Cult., 27, 189-196 (1991); Malik, K. A., Saxena, P. K.,“Regeneration in Phaseolus vulgaris L.—High-Frequency Induction ofDirect Shoot Formation in Intact Seedlings by N-6-Benzylaminopurine andThidiazuron,” 186, 384-389 (3 Feb. 1992); Malik, K. A., Saxena, P. K.,“Somatic Embryogenesis and Shoot Regeneration from Intact Seedlings ofPhaseolus acutifolius A., P. aureus (L.) Wilczek, P. coccineus L., andP. wrightii L.,” Pl. Cell. Rep., 11, 163-168 (3 Apr. 1992); Chavez, J.,et al., “Development of an in vitro culture method for heart shapedembryo in Phaseolus polyanthus,” B.I.C. Invit. Papers 35, 215-216(1992); Munoz-Florez, L. C., et al., “Finding out an efficient techniquefor inducing callus from Phaseolus microspores,” B.I.C. Invit. Papers35, 217-218 (1992); Vaquero, F., et al., “A Method for Long-TermMicropropagation of Phaseolus coccineus L.,” L. Pl. Cell. Rep., 12,395-398 (7-8 May 1993); Lewis, M. E., Bliss, F. A., “Tumor Formation andbeta-Glucuronidase Expression in Phaseolus vulgaris L. Inoculated withAgrobacterium Tumefaciens,” Journal of the American Society forHorticultural Science, 119, 361-366 (2 Mar. 1994); Song, J. Y., et al.,“Effect of auxin on expression of the isopentenyl transferase gene (ipt)in transformed bean (Phaseolus vulgaris L.) single-cell clones inducedby Agrobacterium tumefaciens C58,” J. Plant Physiol. 146, 148-154 (1-2May 1995). It is clear from the literature that the state of the art issuch that these methods of obtaining plants are routinely used and havea very high rate of success. Thus, another aspect of this invention isto provide cells which upon growth and differentiation produce beanplants having the physiological and morphological characteristics ofgarden bean variety PV-803.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, pollen, flowers, seeds, pods, leaves,stems, roots, root tips, anthers, pistils and the like. Means forpreparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture comprising organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and5,977,445 describe certain techniques, the disclosures of which areincorporated herein by reference.

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Such foreignadditional and/or modified genes are referred to herein collectively as“transgenes.” Over the last fifteen to twenty years several methods forproducing transgenic plants have been developed and the presentinvention, in particular embodiments, also relates to transformedversions of the claimed variety or line.

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to, aregulatory element (for example, a promoter). The expression vector maycontain one or more such operably linked gene/regulatory elementcombinations. The vector(s) may be in the form of a plasmid and can beused alone or in combination with other plasmids to provide transformedgarden bean plants using transformation methods as described below toincorporate transgenes into the genetic material of the garden beanplant(s).

Expression Vectors for Garden Bean Transformation: Marker Genes

Expression vectors include at least one genetic marker operably linkedto a regulatory element (a promoter, for example) that allowstransformed cells containing the marker to be either recovered bynegative selection, i.e., inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or an herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene which, when under thecontrol of plant regulatory signals, confers resistance to kanamycin.Fraley, et al., Proc. Natl. Acad. Sci. USA, 80:4803 (1983). Anothercommonly used selectable marker gene is the hygromycinphosphotransferase gene which confers resistance to the antibiotichygromycin. Vanden Elzen, et al., Plant Mol. Biol., 5:299 (1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase and aminoglycoside-3′-adenyltransferase, the bleomycin resistance determinant (Hayford, et al.,Plant Physiol., 86:1216 (1988); Jones, et al., Mol. Gen. Genet., 210:86(1987); Svab, et al., Plant Mol. Biol., 14:197 (1990); Hille, et al.,Plant Mol. Biol., 7:171 (1986)). Other selectable marker genes conferresistance to herbicides such as glyphosate, glufosinate or bromoxynil(Comai, et al., Nature, 317:741-744 (1985); Gordon-Kamm, et al., PlantCell, 2:603-618 (1990); and Stalker, et al., Science, 242:419-423(1988)).

Other selectable marker genes for plant transformation are not ofbacterial origin. These genes include, for example, mouse dihydrofolatereductase, plant 5-enolpyruvylshikimate-3-phosphate synthase and plantacetolactate synthase. Eichholtz, et al., Somatic Cell Mol. Genet.,13:67 (1987); Shah, et al., Science, 233:478 (1986); and Charest, etal., Plant Cell Rep., 8:643 (1990).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase, and chloramphenicol acetyltransferase(Jefferson, R. A., Plant Mol. Biol. Rep., 5:387 (1987); Teeri, et al.,EMBO J., 8:343 (1989); Koncz, et al., Proc. Natl. Acad. Sci. USA, 84:131(1987); DeBlock, et al., EMBO J., 3:1681 (1984)).

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissue are available. However, these in vivomethods for visualizing GUS activity have not proven useful for recoveryof transformed cells because of low sensitivity, high fluorescentbackgrounds and limitations associated with the use of luciferase genesas selectable markers.

A gene encoding Green Fluorescent Protein (GFP) has been utilized as amarker for gene expression in prokaryotic and eukaryotic cells (Chalfie,et al., Science, 263:802 (1994)). GFP and mutants of GFP may be used asscreenable markers.

Expression Vectors for Garden Bean Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are well known in the transformation arts asare other regulatory elements that can be used alone or in combinationwith promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters that initiate transcription only in a certain tissue arereferred to as “tissue-specific.” A “cell-type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may effect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell-type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter that is active under mostenvironmental conditions.

A. Inducible Promoters—An inducible promoter is operably linked to agene for expression in garden bean. Optionally, the inducible promoteris operably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a gene for expression in garden bean. Withan inducible promoter the rate of transcription increases in response toan inducing agent.

Any inducible promoter can be used in the instant invention. See, Ward,et al., Plant Mol. Biol., 22:361-366 (1993). Exemplary induciblepromoters include, but are not limited to, that from the ACEI systemwhich responds to copper (Mett, et al., Proc. Natl. Acad. Sci. USA,90:4567-4571 (1993)); In2 gene from maize which responds tobenzenesulfonamide herbicide safeners (Gatz, et al., Mol. Gen. Genetics,243:32-38 (1994)), or Tet repressor from Tn10 (Gatz, et al., Mol. Gen.Genetics, 227:229-237 (1991)). A particularly preferred induciblepromoter is a promoter that responds to an inducing agent to whichplants do not normally respond. An exemplary inducible promoter is theinducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone (Schena,et al., Proc. Natl. Acad. Sci. USA, 88:0421 (1991)).

B. Constitutive Promoters—A constitutive promoter is operably linked toa gene for expression in garden bean or the constitutive promoter isoperably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a gene for expression in garden bean.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell, et al., Nature, 313:810-812 (1985)) and the promoters from suchgenes as rice actin (McElroy, et al., Plant Cell 2, 163-171 (1990));ubiquitin (Christensen, et al., Plant Mol. Biol., 12:619-632 (1989) andChristensen, et al., Plant Mol. Biol., 18:675-689 (1992)); pEMU (Last,et al., Theor. Appl. Genet., 81:581-588 (1991)); MAS (Velten, et al.,EMBO J., 3:2723-2730 (1984)) and maize H3 histone (Lepetit, et al., Mol.Gen. Genetics, 231:276-285 (1992) and Atanassova, et al., Plant Journal,2 (3):291-300 (1992)). The ALS promoter, Xbal/Ncol fragment 5′ to theBrassica napus ALS3 structural gene (or a nucleotide sequence similarityto said Xbal/Ncol fragment), represents a particularly usefulconstitutive promoter. See, PCT Application WO 96/30530.

C. Tissue-specific or Tissue-preferred Promoters—A tissue-specificpromoter is operably linked to a gene for expression in garden bean.Optionally, the tissue-specific promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in garden bean. Plants transformed with a geneof interest operably linked to a tissue-specific promoter produce theprotein product of the transgene exclusively, or preferentially, in aspecific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promotersuch as that from the phaseolin gene (Murai, et al., Science, 23:476-482(1983) and Sengupta-Gopalan, et al., Proc. Natl. Acad. Sci. USA82:3320-3324 (1985)); a leaf-specific and light-induced promoter such asthat from cab or rubisco (Simpson, et al., EMBO J., 4(11):2723-2729(1985) and Timko, et al., Nature, 318:579-582 (1985)); ananther-specific promoter such as that from LAT52 (Twell, et al., Mol.Gen. Genetics, 217:240-245 (1989)); a pollen-specific promoter such asthat from Zm13 or a microspore-preferred promoter such as that from apg(Twell, et al., Sex. Plant Reprod., 6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall, ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine during protein synthesis and processing where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample, Becker, et al., Plant Mol. Biol., 20:49 (1992); Knox, C., etal., “Structure and Organization of Two Divergent Alpha-Amylase Genesfrom Barley,” Plant Mol. Biol., 9:3-17 (1987); Lerner, et al., PlantPhysiol., 91:124-129 (1989); Fontes, et al., Plant Cell, 3:483-496(1991); Matsuoka, et al., Proc. Natl. Acad. Sci., 88:834 (1991); Gould,et al., J. Cell. Biol., 108:1657 (1989); Creissen, et al., Plant J.,2:129 (1991); Kalderon, et al., Cell, 39:499-509 (1984); Steifel, etal., “Expression of a maize cell wall hydroxyproline-rich glycoproteingene in early leaf and root vascular differentiation,” Plant Cell,2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem., 114:92-6(1981).

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is a garden bean plant. Inanother preferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR and SSR analysis, which identifies the approximate chromosomallocation of the integrated DNA molecule. For exemplary methodologies inthis regard, see Methods in Plant Molecular Biology and Biotechnology,Glick and Thompson Eds., CRC Press, Inc., Boca Raton, 269:284 (1993).Map information concerning chromosomal location is useful forproprietary protection of a subject transgenic plant. If unauthorizedpropagation is undertaken and crosses made with other germplasm, the mapof the integration region can be compared to similar maps for suspectplants, to determine if the latter have a common parentage with thesubject plant. Map comparisons would involve hybridizations, RFLP, PCR,SSR, and sequencing, all of which are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

1. Genes that Confer Resistance to Pests or Disease and that Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with one ormore cloned resistance genes to engineer plants that are resistant tospecific pathogen strains. See, for example, Jones, et al., Science,266:789 (1994) (cloning of the tomato Cf-9 gene for resistance toCladosporium fulvum); Martin, et al., Science, 262:1432 (1993) (tomatoPto gene for resistance to Pseudomonas syringae pv. tomato encodes aprotein kinase); Mindrinos, et al., Cell, 78:1089 (1994) (ArabidopsisRSP2 gene for resistance to Pseudomonas syringae).

B. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser, et al., Gene,48:109 (1986), who disclose the cloning and nucleotide sequence of a Btδ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxin genes canbe purchased from American Type Culture Collection, Manassas, Va., forexample, under ATCC Accession Nos. 40098, 67136, 31995, and 31998.

C. A lectin. See, for example, Van Damme, et al., Plant Molec. Biol.,24:25 (1994), who disclose the nucleotide sequences of several Cliviaminiata mannose-binding lectin genes.

D. A vitamin-binding protein such as avidin. See, PCT Application US93/06487 which teaches the use of avidin and avidin homologues aslarvicides against insect pests.

E. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe, et al., J. Biol. Chem.,262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor); Huub, et al., Plant Molec. Biol., 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I); Sumitani, etal., Biosci. Biotech. Biochem., 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus α-amylase inhibitor).

F. An insect-specific hormone or pheromone such as an ecdysteroid orjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock, et al., Nature, 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

G. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosure of Pratt, et al., Biochem. Biophys. Res. Comm., 163:1243(1989) (an allostatin is identified in Diploptera puntata). See also,U.S. Pat. No. 5,266,317 to Tomalski, et al., which discloses genesencoding insect-specific, paralytic neurotoxins.

H. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang, et al., Gene, 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

I. An enzyme responsible for a hyperaccumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

J. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule, forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase, and a glucanase, whether natural or synthetic. See, PCTApplication WO 93/02197 (Scott, et al.), which discloses the nucleotidesequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also, Kramer, et al., InsectBiochem. Molec. Biol., 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hornworm chitinase, and Kawalleck, et al.,Plant Molec. Biol., 21:673 (1993), who provide the nucleotide sequenceof the parsley ubi4-2 polyubiquitin gene.

K. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella, et al., Plant Molec. Biol., 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess,et al., Plant Physiol., 104:1467 (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

L. A hydrophobic moment peptide. See, PCT Application WO 95/16776, whichdiscloses peptide derivatives of tachyplesin which inhibit fungal plantpathogens, and PCT Application WO 95/18855, which teaches syntheticantimicrobial peptides that confer disease resistance.

M. A membrane permease, a channel former, or a channel blocker. Forexample, see the disclosure of Jaynes, et al., Plant Sci, 89:43 (1993),of heterologous expression of a cecropin-β lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

N. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See, Beachy, et al., Ann. Rev. Phytopathol.,28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virusand tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus, and tobacco mosaic virus. Id.

O. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect.

P. A virus-specific antibody. See, for example, Tavladoraki, et al.,Nature, 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

Q. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo-α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See, Lamb, et al., Bio/Technology,10:1436 (1992).

R. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann, et al., Bio/Technology, 10:305 (1992), have shownthat transgenic plants expressing the barley ribosome-inactivating genehave an increased resistance to fungal disease.

S. Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis-related genes. Briggs, S., “Plant diseaseresistance. Grand unification system theory in sight,” Current Biology,5(2) (1995).

T. Antifungal genes. See, Cornelissen and Melchers, “Strategies forControl of Fungal Diseases with Transgenic Plants,” Plant Physiol.,101:709-712 (1993); and Bushnell, et al., “Genetic Engineering ofDisease Resistance in Cereal,” Can. J. of Plant Path., 20(2):137-149(1998).

2. Genes that Confer Resistance to an Herbicide, for Example:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee, etal., EMBO J., 7:1241 (1988), and Miki, et al., Theor. Appl. Genet.,80:449 (1990), respectively.

B. Glyphosate (resistance conferred by mutant5-enolpyruvlshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus PAT bar genes), and pyridinoxy or phenoxy proprionic acidsand cyclohexones (ACCase inhibitor-encoding genes). See, for example,U.S. Pat. No. 4,940,835 to Shah, et al., which discloses the nucleotidesequence of a form of EPSPS which can confer glyphosate resistance. ADNA molecule encoding a mutant aroA gene can be obtained under ATCCAccession No. 39256, and the nucleotide sequence of the mutant gene isdisclosed in U.S. Pat. No. 4,769,061 to Comai. European PatentApplication No. 0 333 033 to Kumada, et al., and U.S. Pat. No. 4,975,374to Goodman, et al., disclose nucleotide sequences of glutaminesynthetase genes which confer resistance to herbicides such asL-phosphinothricin. See also, Russel, D. R., et al., Plant Cell Report,12:3 165-169 (1993). The nucleotide sequence of aphosphinothricin-acetyl-transferase (PAT) gene is provided in EuropeanApplication No. 0 242 246 to Leemans, et al. DeGreef, et al.,Bio/Technology, 7:61 (1989) describe the production of transgenic plantsthat express chimeric bar genes coding for phosphinothricin acetyltransferase activity. Exemplary of genes conferring resistance tophenoxy proprionic acids and cyclohexones, such as sethoxydim andhaloxyfop, are the Acc1-S1, Acc1-S2, and Acc2-S3 genes described byMarshall, et al., Theor. Appl. Genet., 83:435 (1992).

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibila, et al.,Plant Cell, 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes, et al., Biochem. J.,285:173 (1992).

D. Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants. See, Hattori, et al., “AnAcetohydroxy acid synthase mutant reveals a single site involved inmultiple herbicide resistance,” Mol. Gen. Genet., 246:419-425 (1995).Other genes that confer tolerance to herbicides include a gene encodinga chimeric protein of rat cytochrome P4507A1 and yeast NADPH-cytochromeP450 oxidoreductase (Shiota, et al., “Herbidcide-resistant TobaccoPlants Expressing the Fused Enzyme between Rat Cytochrome P4501A1(CYP1A1) and Yeast NADPH-Cytochrome P450 Oxidoreductase,” PlantPhysiol., 106:17 (1994)), genes for glutathione reductase and superoxidedismutase (Aono, et al., “Paraquat tolerance of transgenic Nicotianatabacum with enhanced activities of glutathione reductase and superoxidedismutase,” Plant Cell Physiol., 36:1687 (1995)), and genes for variousphosphotransferases (Datta, et al., “Herbicide-resistant Indica riceplants from IRRI breeding line IR72 after PEG-mediated transformation ofprotoplants,” Plant Mol. Biol., 20:619 (1992).

E. Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306, 6,282,837,5,767,373, and International Publication WO 01/12825.

3. Genes that Confer or Contribute to a Value-Added Trait, Such as:

A. Delayed and attenuated symptoms to Bean Golden Mosaic Geminivirus(BGMV), for example, by transforming a plant with antisens genes fromthe Brazilian BGMV. See, Arago, et al., Molecular Breeding, 4:6, 491-499(1998).

B. Increased the bean content in Methionine by introducing a transgenecoding for a Methionine rich storage albumin (2S-albumin) from theBrazil nut as described in Arago, et al., Genetics and MolecularBiology, 22:3, 445-449 (1999).

Methods for Garden Bean Transformation

Numerous methods for plant transformation have been developed includingbiological and physical plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants,” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson Eds., CRC Press, Inc., Boca Raton, pp. 67-88 (1993). Inaddition, expression vectors and in-vitro culture methods for plant cellor tissue transformation and regeneration of plants are available. See,for example, Gruber, et al., “Vectors for Plant Transformation” inMethods in Plant Molecular Biology and Biotechnology, Glick and ThompsonEds., CRC Press, Inc., Boca Raton, pp. 89-119 (1993).

A. Agrobacterium-mediated Transformation—One method for introducing anexpression vector into plants is based on the natural transformationsystem of Agrobacterium. See, for example, Horsch, et al., Science,227:1229 (1985); Diant, et al., Molecular Breeding, 3:1, 75-86 (1997).A. tumefaciens and A. rhizogenes are plant pathogenic soil bacteriawhich genetically transform plant cells. The Ti and Ri plasmids of A.tumefaciens and A. rhizogenes, respectively, carry genes responsible forgenetic transformation of the plant. See, for example, Kado, C. I.,Crit. Rev. Plant Sci., 10:1 (1991). Descriptions of Agrobacterium vectorsystems and methods for Agrobacterium-mediated gene transfer areprovided by Gruber, et al., supra, Miki, et al., supra, and Moloney, etal., Plant Cell Reports, 8:238 (1989). See also, U.S. Pat. No. 5,591,616issued Jan. 7, 1997.

B. Direct Gene Transfer—Despite the fact the host range forAgrobacterium-mediated transformation is broad, some cereal or vegetablecrop species and gymnosperms have generally been recalcitrant to thismode of gene transfer, even though some success has been achieved inrice and corn. Hiei, et al., The Plant Journal, 6:271-282 (1994) andU.S. Pat. No. 5,591,616 issued Jan. 7, 1997. Several methods of planttransformation, collectively referred to as direct gene transfer, havebeen developed as an alternative to Agrobacterium-mediatedtransformation. A generally applicable method of plant transformation ismicroprojectile-mediated transformation where DNA is carried on thesurface of microprojectiles measuring 1 to 4 microns. The expressionvector is introduced into plant tissues with a biolistic device thataccelerates the microprojectiles to speeds of 300 to 600 m/s which issufficient to penetrate plant cell walls and membranes. Russell, D. R.,et al., Pl. Cell. Rep., 12, 165-169 (3 Jan. 1993); Aragao, F. J. L., etal., Plant Mol. Biol., 20, 357-359 (2 Oct. 1992); Aragao, Theor. Appl.Genet., 93:142-150 (1996); Kim, J.; Minamikawa, T., Plant Science,117:131-138 (1996); Sanford, et al., Part. Sci. Technol., 5:27 (1987);Sanford, J. C., Trends Biotech., 6:299 (1988); Klein, et al., Bio/Tech.,6:559-563 (1988); Sanford, J. C., Physiol Plant, 7:206 (1990); Klein, etal., Biotechnology, 10:268 (1992).

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang, et al., Bio/Technology, 9:996 (1991).Alternatively, liposome and spheroplast fusion have been used tointroduce expression vectors into plants. Deshayes, et al., EMBO J.,4:2731 (1985); Christou, et al., Proc Natl. Acad. Sci. USA, 84:3962(1987). Direct uptake of DNA into protoplasts using CaCl₂ precipitation,polyvinyl alcohol or poly-L-ornithine have also been reported. Hain, etal., Mol. Gen. Genet., 199:161 (1985) and Draper, et al., Plant CellPhysiol., 23:451 (1982). Electroporation of protoplasts and whole cellsand tissues have also been described Saker, M. and Kuhne, T., BiologiaPlantarum, 40(4):507-514 (1997/98); D'Halluin, et al., Plant Cell,4:1495-1505 (1992); and Spencer, et al., Plant Mol. Biol., 24:51-61(1994)).

Following transformation of garden bean target tissues, expression ofthe above-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic variety. The transgenic variety could then becrossed with another (non-transformed or transformed) variety in orderto produce a new transgenic variety. Alternatively, a genetic trait thathas been engineered into a particular garden bean line using theforegoing transformation techniques could be moved into another lineusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite variety into anelite variety, or from a variety containing a foreign gene in its genomeinto a variety or varieties that do not contain that gene. As usedherein, “crossing” can refer to a simple X by Y cross or the process ofbackcrossing depending on the context.

Backcrossing

When the term garden bean plant, cultivar or bean line are used in thecontext of the present invention, this also includes cultivars where oneor more desired traits has been introduced through backcrossing methods,whether such trait is a naturally occurring one or a transgenic one.Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the line. The term “backcrossing” asused herein refers to the repeated crossing of a hybrid progeny back tothe recurrent parent, i.e., backcrossing one, two, three, four, five,six, seven, eight, nine, or more times to the recurrent parent. Theparental bean plant which contributes the gene for the desiredcharacteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental bean plant to which the gene or genes from the nonrecurrentparent are transferred is known as the recurrent parent as it is usedfor several rounds in the backcrossing protocol.

In a typical backcross protocol, the original cultivar of interest(recurrent parent) is crossed to a second line (nonrecurrent parent)that carries the gene or genes of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a garden bean plantis obtained wherein essentially all of the desired morphological andphysiological characteristics of the recurrent parent are recovered inthe converted plant, generally determined at a 5% significance levelwhen grown in the same environmental condition, in addition to the geneor genes transferred from the nonrecurrent parent. It has to be notedthat some, one, two, three, or more, self-pollination and growing ofpopulation might be included between two successive backcrosses. Indeed,an appropriate selection in the population produced by theself-pollination, i.e., selection for the desired trait andphysiological and morphological characteristics of the recurrent parentmight be equivalent to one, two or even three, additional backcrosses ina continuous series without rigorous selection, saving time, money andeffort to the breeder. A non limiting example of such a protocol wouldbe the following: (a) the first generation F₁ produced by the cross ofthe recurrent parent A by the donor parent B is backcrossed to parent A;(b) selection is practiced for the plants having the desired trait ofparent B; (c) selected plants are self-pollinated to produce apopulation of plants where selection is practiced for the plants havingthe desired trait of parent B and the physiological and morphologicalcharacteristics of parent A; (d) the selected plants are backcrossedone, two, three, four, five, six, seven, eight, nine, or more times toparent A to produce selected backcross progeny plants comprising thedesired trait of parent B and the physiological and morphologicalcharacteristics of parent A. Step (c) may or may not be repeated andincluded between the backcrosses of step (d).

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalline. To accomplish this, a gene or genes of the recurrent cultivar ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphological,constitution of the original line. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross, one ofthe major purposes is to add some commercially desirable, agronomicalimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a single geneand dominant allele, multiple genes and recessive allele(s) may also betransferred and therefore, backcross breeding is by no means restrictedto character(s) governed by one or a few genes. In fact the number ofgenes might be less important than the identification of thecharacter(s) in the segregating population. In this instance it may thenbe necessary to introduce a test of the progeny to determine if thedesired characteristic(s) has been successfully transferred. Such testsencompass visual inspection, simple crossing but also follow up of thecharacteristic(s) through genetically associated markers and molecularassisted breeding tools. For example, selection of progeny containingthe transferred trait is done by direct selection, visual inspection fora trait associated with a dominant allele, while the selection ofprogeny for a trait that is transferred via a recessive allele requiresselfing the progeny to determine which plant carries the recessiveallele(s).

Many single gene traits have been identified that are not regularlyselected for in the development of a new line but that can be improvedby backcrossing techniques. Single gene traits may or may not betransgenic. Examples of these traits include, but are not limited to,herbicide resistance (such as bar or pat genes), resistance forbacterial, fungal, or viral disease (such as gene I used for BCMVresistance), insect resistance, enhanced nutritional quality (such as 2salbumine gene), industrial usage, agronomic qualities (such as the“persistent green gene”), yield stability, and yield enhancement. Thesegenes are generally inherited through the nucleus. Some other singlegene traits are described in U.S. Pat. Nos. 5,777,196, 5,948,957, and5,969,212, the disclosures of which are specifically hereby incorporatedby reference.

In 1981 the backcross method of breeding accounted for 17% of the totalbreeding effort for inbred corn line development in the United States,according to, Hallauer, A. R., et al., “Corn Breeding,” Corn and CornImprovement, No. 18, pp. 463-481 (1988).

The backcross breeding method provides a precise way of improvingvarieties that excel in a large number of attributes but are deficientin a few characteristics. (Page 150 of the Pr. R. W. Allard's 1960 book,Principles of Plant Breeding, published by John Wiley & Sons, Inc.) Themethod makes use of a series of backcrosses to the variety to beimproved during which the character or the characters in whichimprovement is sought is maintained by selection. At the end of thebackcrossing the gene or genes being transferred unlike all other genes,will be heterozygous. Selfing after the last backcross produceshomozygosity for this gene pair(s) and, coupled with selection, willresult in a variety with exactly the adaptation, yielding ability, andquality characteristics of the recurrent parent but superior to thatparent in the particular characteristic(s) for which the improvementprogram was undertaken. Therefore, this method provides the plantbreeder with a high degree of genetic control of his work.

When using a transgene, the trait is generally not incorporated intoeach newly developed line/cultivar such as bean variety PV-803 by directtransformation. Rather, the more typical method used by breeders ofordinary skill in the art to incorporate the transgene is to take a linealready carrying the transgene and to use such line as a donor line totransfer the transgene into the newly developed line. The same wouldapply for a naturally occurring trait. The backcross breeding processcomprises the following steps: (a) crossing bean variety PV-803 plantswith plants of another variety that comprise the desired trait(s); (b)selecting the F₁ progeny plants that have the desired trait(s); (c)crossing the selected F₁ progeny plants with bean variety PV-803 plantsto produce backcross progeny plants; (d) selecting for backcross progenyplants that have the desired trait(s) and physiological andmorphological characteristics of bean variety PV-803 to produce selectedbackcross progeny plants; and (e) repeating steps (c) and (d) one, two,three, four, five six, seven, eight, nine, or more times in successionto produce selected, second, third, fourth, fifth, sixth, seventh,eighth, ninth, or higher backcross progeny plants that comprise thedesired trait(s) and the physiological and morphological characteristicsof bean variety PV-803 as determined in Table 1 at a 5% significancelevel when grown in the same environmental conditions. The bean plantsproduced by the methods are also part of the invention.

The backcross method is scientifically exact because the morphologicaland agricultural features of the improved variety could be described inadvance and because the same variety could, if it were desired, be breda second time by retracing the same steps (Briggs, “Breeding wheatsresistant to bunt by the backcross method,” Jour. Amer. Soc. Agron.,22:289-244 (1930)).

Backcrossing is a powerful mechanism for achieving homozygosity and anypopulation obtained by backcrossing must rapidly converge on thegenotype of the recurrent parent. When backcrossing is made the basis ofa plant breeding program, the genotype of the recurrent parent will bemodified only with regards to genes being transferred, which aremaintained in the population by selection.

TABLES

In Table 2, the traits and characteristics of garden bean variety PV-803are compared to those of commercial variety Valentino (U.S. PlantVariety Protection No. 200600097). The data was collected during2009-2012 from several locations in the United States and Europe.

The first column shows the variety name. The second column shows thelocation of testing. The third column shows the pod diameter inmillimeters. The fourth column shows the pod color on a scale of 1 to 9(1 being very pale and 9 being the darkest).

TABLE 2 Pod Variety Location Diameter (mm) Pod Color Valentino Florida2009 9.5 8 PV-803 Florida 2009 9.5 9 Valentino Netherlands 2009 9 PV-803Netherlands 2009 10 8 Valentino Florida 2010 9 8 PV-803 Florida 2010 9.59 Valentino Wisconsin 2011 10 7 PV-803 Wisconsin 2011 10 9 ValentinoFlorida 2012 10 8 PV-803 Florida 2012 10.5 8.5

As shown in Table 2, garden bean variety PV-803 has an overall slightlylarger pod diameter when compared to Valentino. In addition, the podcolor of PV-803 is overall slightly darker than Valentino.

DEPOSIT INFORMATION

A deposit of the Pop Vriend Research BV proprietary GARDEN BEAN VARIETYPV-803 disclosed above and recited in the appended claims has been madewith the American Type Culture Collection (ATCC), 10801 UniversityBoulevard, Manassas, Va. 20110. The date of deposit was Nov. 1, 2012.The deposit of 2,500 seeds was taken from the same deposit maintained byPop Vriend Research BV since prior to the filing date of thisapplication. All restrictions will be irrevocably removed upon grantingof a patent, and the deposit is intended to meet all of the requirementsof 37 C.F.R. §§1.801-1.809. The ATCC Accession Number is PTA-13288. Thedeposit will be maintained in the depository for a period of thirtyyears, or five years after the last request, or for the enforceable lifeof the patent, whichever is longer, and will be replaced as necessaryduring that period.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions and sub-combinations as are within their truespirit and scope.

What is claimed is:
 1. A seed of bean variety designated PV-803, whereina representative sample of seed of said variety has been deposited underPTA-13288.
 2. A bean plant, or a part thereof, produced by growing theseed of claim
 1. 3. A tissue culture of regenerable cells produced fromthe plant of claim 2, wherein said cells of the tissue culture areproduced from a plant part selected from the group consisting ofembryos, meristematic cells, leaves, pollen, root, root tips, stems,anther, pistils, pods, flowers, and seeds.
 4. A bean plant regeneratedfrom the tissue culture of claim 3, said plant having all of themorphological and physiological characteristics of bean variety PV-803,wherein a representative sample of seed of said variety has beendeposited under PTA-13288.
 5. A method for producing a bean seedcomprising crossing a first parent bean plant with a second parent beanplant and harvesting the resultant hybrid bean seed, wherein said firstparent bean plant or second parent bean plant is the bean plant of claim2.
 6. A hybrid bean seed produced by the method of claim
 5. 7. A methodfor producing an herbicide resistant bean plant comprising transformingthe bean plant of claim 2 with a transgene that confers herbicideresistance to an herbicide selected from the group consisting ofimidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine, and benzonitrile.
 8. An herbicideresistant bean plant, or a part thereof, produced by the method of claim7.
 9. A method for producing an insect resistant bean plant comprisingtransforming the bean plant of claim 2 with a transgene that confersinsect resistance.
 10. An insect resistant bean plant, or a partthereof, produced by the method of claim
 9. 11. A method for producing adisease resistant bean plant comprising transforming the bean plant ofclaim 2 with a transgene that confers disease resistance.
 12. A diseaseresistant bean plant, or a part thereof, produced by the method of claim11.
 13. A method of introducing a desired trait into bean variety PV-803comprising: (a) crossing a bean variety PV-803 plant grown from beanvariety PV-803 seed, wherein a representative sample of seed of saidvariety has been deposited under PTA-13288, with another bean plant thatcomprises a desired trait to produce F₁ progeny plants, wherein thedesired trait is selected from the group consisting of insectresistance, disease resistance, water stress tolerance, heat tolerance,improved shelf life, and improved nutritional quality; (b) selecting oneor more progeny plants that have the desired trait; (c) backcrossing theselected progeny plants with bean variety PV-803 to produce backcrossprogeny plants; (d) selecting for backcross progeny plants that have thedesired trait and all of the physiological and morphologicalcharacteristics of bean variety PV-803 listed in Table 1; and (e)repeating steps (c) and (d) three or more times in succession to produceselected fourth or higher backcross progeny plants that comprise thedesired trait and all of the physiological and morphologicalcharacteristics of bean variety PV-803 listed in Table
 1. 14. A beanplant produced by the method of claim 13, wherein the plant has thedesired trait and all of the physiological and morphologicalcharacteristics of bean variety PV-803 listed in Table
 1. 15. The beanplant of claim 14, wherein the desired trait is herbicide resistance andthe resistance is conferred to an herbicide selected from the groupconsisting of imidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine, and benzonitrile.
 16. The bean plant ofclaim 14, wherein the desired trait is insect resistance and the insectresistance is conferred by a transgene encoding a Bacillus thuringiensisendotoxin.
 17. The bean plant of claim 14, wherein the desired trait isselected from the group consisting of insect resistance, diseaseresistance, water stress tolerance, heat tolerance, improved shelf life,and improved nutritional quality.
 18. A method of producing a commodityplant product, comprising obtaining the plant of claim 2, or a partthereof, and producing the commodity plant product from said plant orplant part thereof, wherein said commodity plant product is selectedfrom the group consisting of protein concentrate, protein isolate,garden bean hulls, vegetable garden bean, meal, flour and oil.